不同种属真皮成纤维细胞在不同血清浓度培养基中生长的比较观察

Proliferation of dermal fibroblasts from human,Yorkshire pigsor SD rats cultured in DMEM with different concentrations of serum

目的 :探讨作为皮肤组织工程种子细胞的不同种属动物成纤维细胞在不同培养基中的生长状态 ,以获得最佳生长状态的种子细胞。方法 :分别配制体积分数为 10 0、50、2 0ml/L血清和 2 0ml/L血清 +牛垂体提取物 (BPE ,0 .0 2 5g/L) 4种培养基来培养人、York猪、SD鼠的真皮成纤维细胞 ,通过MTT比色和BrdU检测观察细胞的生长状态。结果 :成纤维细胞在 4种培养基中的生长状态有所不同 :人成纤维细胞在 10 0、50ml/L血清和 2 0ml/L血清 +BPE的培养基中生长良好 ,York猪成纤维细胞在 50ml/L血清和 2 0ml/L血清 +BPE的培养基中生长良好 ,SD鼠成纤维细胞只有在 2 0ml/L血清 +BPE的培养基中生长状态好 ,增殖快。结论 :不同浓度血清的培养液对真皮成纤维细胞的生长都有影响 ,血清体积分数为 2 0ml/L(含BPE ,0 .0 2 5g/L)的培养液对3种真皮成纤维的生长都适合 ,血清体积分数为 10 0ml/L和 50ml/L的培养液也分别适合于人。

Objective: To establish an optimized culture condition f or the growth of dermal fibroblasts. Methods: Human, Yorks hire pig or SD rat fibroblasts were cultured in DMEM with fetal boven serum(P BS) at the concentrations(ml/L) of 100 (A), 50 (B), 20 (C) or 20 supplemente d with 0.025 g/L boven pituitary extract (BPE)(D). The proliferation of the cell s were examined by microscopic observation,MTT assay and BrdU analysis. Results: Human dermal fibroblasts grew better in the medium with FBS at 100 ml/L, 50 ml/L and 20 ml/L supplemented with BPE, Yorkshire pig dermal fibroblasts did in medium with FBS at 50 ml/L and 20 ml/L supplemented with BPE . SD rat dermal fibroblasts did in the medium with 20 ml/L FBS supplemented wi th BPE. Conclusion:The favarable serum concentration in me dium for human, Yorkshire pigs and SD rats dermal fibroblasts was different,but the medium with 20 ml/L FBS supplement with BPE is suitable for all 3 kinds of f ibroblasts.